After 50-fold overdigestion with ScaI, more than 80% of the DNA fragments can be ligated and more than 90% of these can be recut

Methylation Effects

Dam: never overlaps — no effect

Dcm: never overlaps — no effect

CpG: never overlaps — no effect

EcoKI: never overlaps — no effect

EcoBI: may overlap — blocked

Digestion of Agarose-embedded DNA

Minimum 20units of the enzyme are required for complete digestion of 1μg of agarose-embedded lambda DNA in 16hours

Note:

Conditions of low salt, high enzyme concentration, glycerol concentration > 5% or pH> 8.0 may result in star activity. Supercoiled plasmids may require up to 20-fold more ScaI for complete digestion than linear DNA.